Novel Biodegradable Nanoadjuvants for Immunotherapy of Pancreatic Cancer

描述（由申请人提供）：目前的医学疗法未能有效治疗胰腺癌。由于平均存活率仍低于4%，迫切需要研究替代治疗方法。该项目的总体目标是开发一种新的纳米级疫苗递送系统，该系统将导致对胰腺癌的强烈抗肿瘤反应。肿瘤相关抗原Mucin-1在胰腺癌中过度表达和差异糖基化。我们将设计疫苗制剂，将粘蛋白-1封装到具有免疫调节能力的新型两亲性聚酸酐共聚物系统的纳米球中。具体目标是：（SA 1）通过由1，6-双-（对-羧基苯氧基）己烷（CPH）和1，8-双-（对-羧基苯氧基）-3，6-二氧杂辛烷（CPTEG）组成的生物可降解的两亲性聚酸酐纳米球测定树突细胞（DC）的细胞毒性和活化的体外研究;和（SA 2）使用鼠模型测定其免疫原性和免疫调节能力的装载粘蛋白-1的纳米球的体内研究。我们的方法结合了抗原呈递细胞（APC）的摄取效率，这些细胞的活化和迁移，以及诱导适当的细胞介导的免疫（即抗原特异性T细胞反应）的阐明。由于颗粒大小对有效的吞噬作用起着重要作用，SA 1将涉及通过不同技术制造微/纳米球：固体/油/油双乳液，低温芳香化和纳米沉淀。用鼠骨髓来源的树突状细胞培养微/纳米球将研究在MHC I和MHC II分子的背景下DC的细胞毒性和有效活化。在确定哪种CPTEG：CPH制剂诱导Th 1细胞因子后，将在鼠模型上评价这些细胞因子，以鉴定将促进诱导抗肿瘤免疫的制剂。免疫应答途径和纳米球佐剂的免疫调节能力（即，Th 1 vs. Th 2）将通过细胞因子谱和产生的抗体进行评价。根据获得的结果，将在野生型（C57 BL/6）小鼠中评价抗肿瘤应答。该项目的预期成果是确定CPTEG：CPH微/纳米球制剂，用于在转基因小鼠模型中进行进一步测试，以评价耐受性和自身免疫背景下的抗肿瘤免疫力，这将导致基于疫苗的免疫疗法。本案无关该技术的开发和应用将促进抗肿瘤疫苗的合理设计，其具有适当调节免疫应答以产生保护性免疫的能力，并导致将彻底改变抗肿瘤免疫疗法的技术。

DESCRIPTION (provided by applicant): Current medical therapies have failed to treat pancreatic cancer effectively. There is an urgent need to investigate alternative treatments as the average survival rate remains less than 4%. The overall objective of this project is to develop a novel nanoscale vaccine delivery system that will lead to strong anti-tumor responses against pancreatic cancer. The tumor-associated antigen Mucin-1 is over expressed and differentially glycosylated by pancreatic adenocarcinomas. We will design vaccine formulations that encapsulate Mucin-1 into nanospheres of a novel amphiphilic polyanhydride copolymer system with immunomodulatory capabilities. The specific aims are: (SA1) In vitro studies to determine cytotoxicity and activation of dendritic cells (DCs) by biodegradable amphiphilic polyanhydride nanospheres composed of 1,6-bis-(p-carboxyphenoxy)hexane (CPH) and 1,8-bis-(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG); and (SA2) In vivo studies of Mucin-1 loaded nanospheres using murine models to determine their immunogenicity and immunomodulatory capabilities. Our approach combines the elucidation of the uptake efficiency of antigen presenting cells (APCs), the activation and migration of these cells, and the induction of appropriate cell-mediated immunity (i.e. antigen-specific T cell responses). Since particle size plays a major role for efficient phagocytosis, SA1 will involve the fabrication of micro/nanospheres by different techniques: solid/oil/oil double emulsion, cryogenic aromization, and nanoprecipitation. Culture of the micro/nanospheres with murine bone marrow derived dendritic cells will investigate the cytotoxicity and efficient activation of DCs in the context of MHC I and MHC II molecules. After determination of which CPTEG:CPH formulation induce Th1 cytokines, these will be evaluated on murine models to identify formulations that will facilitate the induction of anti-tumor immunity. The immune response pathway and the immunomodulatory capabilities of the nanosphere adjuvants (i.e., Th1 vs. Th2) will be evaluated by means of the cytokine profiles and antibodies produced. Based on the results obtained, an anti-tumor response will be evaluated in wild type (C57BL/6) mice. The expected outcome of this project is the determination of CPTEG:CPH micro/nanosphere formulations for further testing in transgenic murine models to evaluate anti-tumor immunity in the context of tolerance and auto-immunity, which will lead to vaccine-based immunotherapies. Relevance. Development and application of this technology will facilitate the rational design of anti-tumor vaccines with the ability to appropriately modulate the immune response to develop protective immunity and lead to technologies that will revolutionize anti-tumor immunotherapies.